Our previous reports showed that exposure to the neonicotinoid pesticide clothianidin (CLO) at a no-observed-adverse-effect-level (NOAEL) dose during fetal development and lactation in mice led to higher rates of maternal neglect and infanticide. Although the demonstrated association between decreased oxytocin secretion and decreased maternal parenting behavior implies a link to declining oxytocin levels, no evidence has yet emerged in CLO to clearly establish such an association. This study investigated the effects of CLO on maternal behavior and oxytocin in C57BL/6N mice exposed during pregnancy and lactation (F0 mothers) as well as in their adult female offspring (F1 mothers). The effects were assessed using nest building assays during pregnancy and pup retrieval assessment after delivery. The results showed a decrease in oxytocin secretion and a marked decrease in pup retrieval behavior among the F0 mothers in the CLO exposure group compared to those in the control group. Their offspring, the F1 mothers, showed significantly lower nest-building scores during pregnancy. In conclusion, this study is the first to examine the potential mechanisms by which CLO exposure in mothers at the NOAEL dose during pregnancy and lactation results in reduced plasma oxytocin levels, subsequently leading to a decline in maternal behaviors such as pup retrieval. Furthermore, these effects may impair maternal behaviors in the next generation, when the offspring mice become mothers.